꿀벌에 대한 dsRNA의 급성섭식독성 평가

임혜송,정영준,김일룡,김진,유성민,김반니,이중로,최원균 한국환경농학회 2017 한국환경농학회지 Vol.36 No.4

본 연구는 최근 RNAi 기반 LMO의 연구․개발이 활발히진행됨에 따라 향후 이러한 기술을 이용한 LMO의 유해성및 자연생태계 위해성평가가 필요할 때 실험실 수준에서dsRNA를 대량으로 발현시키는 시스템을 확립하고, 수분(화분)매개 곤충인 꿀벌을 대상으로 유해성평가 시험을 수행하는 방법을 제시하고자 하였다. L4440 vector에 Snf7과 GFP 유전자를 클로닝한 plasmid를 HT115 (DE3) 대장균에 형질전환한 후 온도, 배양시간, IPTG 농도를 각기 다르게 하여최적의 발현조건을 탐색한 결과 37℃, 0.4 mM IPTG, 4시간의 배양시간에서 가장 많은 양의 dsRNA가 발현됨을 확인하였다. 국내․외 제시된 꿀벌 위해성평가 가이드라인을 바탕으로 대장균에서 분리한 dsRNA를 꿀벌 성충에 급성섭식으로처리한 결과 생사율과 일반중독증상에서 차이를 보이지 않는것으로 보아 대장균으로부터 분리한 Snf7 dsRNA와 GFP dsRNA는 꿀벌 성충에 유해하지 않음을 알 수 있었다. 본 연구를 통해 dsRNA 물질의 유해성평가 및 자연생태계 위해성평가를 위한 대량 추출 방법과 위해성평가 대상종의 사육 및물질 처리 방법을 확립하여 향후 이뤄질 dsRNA의 꿀벌 위해성평가에 활용될 것으로 사료된다. BACKGROUND:RNAinterference (RNAi) eliminates or decreases gene expression by disrupting the target mRNA or by interfering with translation. Recently, RNAi technique was applied to generate new crop traits which provide protection against pests. To establish the environmental risk assessment protocol of RNAi LMO in lab scale,we developed dsRNAexpression systemusing E. coli and tested acute oral toxicity assay to honey. METHOD AND RESULTS: The dsRNA expression vector, L4440, was chosen and cloned 240 bp of Snf7 and GFP gene fragment. To develop the maximum dsRNA induction condition in E. coli, we tested induction time, temperature and IPTGconcentration inmedia. To estimate the risk assessment of dsRNA to honey bee, it has been selected and culturedwith dsRNAsupplement for 48 hours according to OECD guideline. As a result, the optimum condition of dsRNA induction was 37℃, 4 hours and 0.4 mM IPTG concentration and the difference between Snf7 andGFP dsRNAmolecules fromE. coliwas not significant in survival and behavior to honey bee. Furthermore, blast search results indicated that effective match of predicted dsRNA fragments were not existed in honey bee genome. CONCLUSION: In this study,we developed and tested the acute oral toxicity of dsRNA using E. coli expression system to honey bee.

젠더의 모방과 전복: 클로드 카엔의 자화상 사진

임혜송 서양미술사학회 2019 서양미술사학회논문집 Vol.50 No.-

This study examines imitation and subversion of gender in Claude Cahun’s self-portraits by representing lesbian subjects. Masquerade of gender codes and dissolution of boundaries visualize gestures of gender violations in the 1920’s lesbian literature, challenging phallus-centered authority and distorted views on homosexuality by the medium of female body. Metaphorizing her bisexual self-identity through ‘cross-dressing’, Cahun criticizes and imitates paternal power. Parodying the self, a lesbian artist with Jewish identity, concretizes fashion politics for subjective female power. Imitation of femininity, weakening the boundary between reality and artificiality, embodies lesbian subjects’s desire and femininity’s artificial substance. Gender binding and psychological concealment reveal her critical gaze on patriarchal cultural discourse in Western Christianity. Performing exaggerated femininity and victims of gender oppression with dolls and masks, she attempts ‘re-visualizing’ female gender and ‘defensive’ concealment of the existence. Cahun raised the question of photographic nature of reality representation by subverting the boundary between reason and irrationality through political, aesthetic performances of lesbian divisive gender and self-extinction. While projecting oppressed lesbian subjects’ desire and bisexual identity, irrational metaphors of surrealist aesthetics and gender binding are interpreted as Cahun’s psychological rhetoric to conceal and minimize the self-existence. 본 연구는 클로드 카엔의 자화상 사진에 나타난 젠더의 모방과 전복성을 레즈비언 주체의 재현 전략을 통해 살펴본다. 1920년대 레즈비언 문학 속에 나타난, 현대성의 상징으로 대표되는 신여성의 젠더위반의 제스처는 젠더 코드의 가장과 경계의 해체로 가시화된다. 당대 레즈비언 지식인의 부계 권력의비판과 모방은 ‘복장 도착’으로서 양성적 자아 정체성을 과시하고 은유한다. 특히 서구 기독교 문화 속에서 인종적 타자로 배척당해온 유대 가계의 정체성과 레즈비언 예술가로서의 자아를 패러디하는 카엔의 자아 재현 전략은 주체적 여성 권력에 대한 패션의 정치학을 통해 가시화된다. 카엔의 현실과 인공의 경계를 약화시키는 ‘여성스러움의 모방’은 레즈비언 주체의 욕망과 여성성의 인공적 현실을 암시하는 양가적 의미를 갖는다. 마스크와 인형으로 가장한 연극적 자화상에서 카엔은 수동적인 젠더 억압의희생물, 여성성의 과장된 이미지로 퍼포먼스 함으로써 여성 젠더의 ‘재가시화’를 시도한다. 서구 기독교의 가부장적 문화 담론에 대한 카엔의 비판적 시선은 성적, 인종적 타자로서의 자아 정체성 재현에서젠더 묶기와 은폐의 심리적 가장으로 요약할 수 있다. 작가는 이성과 비이성의 경계를 전복시킴으로써사실 재현이라는 사진적 속성에 의문을 제기하였고, 이것은 레즈비언의 분열적 젠더와 자아 소멸의 정치적, 미학적 퍼포먼스로써 가시화되었다. 카엔은 여성의 몸을 매개로 한 젠더의 가장과 허구적 페르소나의 모방을 통해 남근 중심의 권위와 동성애에 대한 왜곡된 시각에 이의를 제기한다. 초현실주의 미학의 비이성적 환유와 젠더 묶기는 억압된 레즈비언 주체의 욕망과 양성적 젠더 정체성을 투사하는 동시에 나치의 위협으로부터 자아 존재를 은폐하고 최소화시키려는 카엔의 심리적 수사로 해석된다.

dsRNA의 꿀벌 급성섭식독성평가

임혜송(Hye Song Lim),정영준(Young Jun Jung),김일룡(Il Ryong Kim),이중로(Jung Ro Lee),최원균(Wonkyun Choi) 환경독성보건학회 2017 한국독성학회 심포지움 및 학술발표회 Vol.2017 No.10

Application of simple and massive purification system of dsRNA in vivo for acute toxicity to Daphnia magna

최원균,임혜송,김진,류성민,이정로 한국곤충학회 2018 Entomological Research Vol.48 No.6

The RNA interference (RNAi) has been considered as an important genetic tool and applied to develop a new living modified (LM) crop trait which is an improvement of nutrient quality or pest management. The RNAi of DvSnf7 has been used for resistance to LM maize and the Western Corn Rootworm which is a major agricultural pest for the US Corn Belt. Most of the environmental risk assessments (ERA) of double strand RNA (dsRNA) have been performed using in vitro transcript products, and not in vivo expressed product. A large amount of dsRNA was required for the acute toxicity assay of water fleas. Therefore development of massive dsRNA purification techniques is critical. Daphnia, a freshwater microcrustacean, is a model organism for studying cellular and molecular mechanism involved in life history traits and ecotoxicology. In this study, we established the massive dsRNA purification method using Escherichia coli and implemented acute toxicity assays to Daphnia magna. As a result, the present RNase A and DNase I, dsRNA was efficiently purified without any special techniques or equipment. Even though purified dsRNA existed during the acute toxicity test, lethality or abnormal behavior were not observed in D. magna. These results indicated that GFP and DvSnf7 dsRNA were not significantly affected to D. magna due to their lack of sequence matching in its genome. The purification method of dsRNA and the acute toxicity assay of water fleas using purified dsRNA would be suitable for the toxicological studies of LMOs to aquatic non‐target organisms.

국내 LMO 자연환경 모니터링을 위한 11개 LM 옥수수의 동시검출기법 개발

신수영,임혜송,설민아,정영준,김일룡,송해룡,이중로,최원균 한국식물생명공학회 2016 JOURNAL OF PLANT BIOTECHNOLOGY Vol.43 No.4

With the increasing development and commercial use of genetically modified maize, it is essential to develop an appropriate method for detection of individual LMO (Living modified organism) events for monitoring the samples. In South Korea, commercial planting and accidental or unintentional releases of LMOs into the environment were not approved. In this study, to increase the efficiency of LMO detection, we developed simultaneous detection methods for 11 LM maize events. This multiplex PCR detection method is economical, as it saves time, cost and labor. We developed 11 individual LM maize events, and applied 4 multiplex PCR sets to the LM maize samples. These results are confirmed by applying the multiplex analysis of LMO environmental monitoring from 2012 to 2014, which represents the unintentionally released LM maize samples. The data were correlated with event specific PCR results. Our results indicate that the multiplex PCR method developed is suitable for detection of LM maize in LMO monitoring. 유전자변형 옥수수의 개발과 상업적 이용이 증가하면서 유전자변형 옥수수 의심시료의 LMO 이벤트를 확인 할 수 있는 적합한 방법 개발이 필요하다. 국내에서는 상업적 재배와 자연생태계의 의도적·비의도적 LMO의 유출이 허용되고있지 않다. 본 연구에서는 국내 승인된 11개의 LM 옥수수 이벤트를 동시에 검출할 수 있는 동시검출기법을 개발하였다. 이 방법은 시간과 비용, 노동력을 절감할 수 있는 효율적인방법으로 4종의 PCR set로 개발하였다. 2012년부터 2014년까지 국립환경과학원 및 국립생태원에서 실시한 LMO 자연환경모니터링 의심시료를 이용하여 동시검출기법의 효율성을 검증하였다. 이러한 결과를 바탕으로 본 연구의 결과는 LMO 모니터링 시료의 분석에 적합한 방법임을 알 수 있었다.

Investigation of Agrobacterium-mediated Transient dsRNA Expression in Tobacco

최원균,임혜송,서한규,김동욱 한국하천호수학회 2019 생태와 환경 Vol.52 No.4

The Agrobacterium tumefaciens mediated gene transfer is widely used to generate genetic transformation of plants and transient assay of temporal exogenous gene expression. Syringe infiltration system into tobacco (Nicotiana benthamiana) leaves is a powerful tool for transient expression of target protein to study protein localization, protein-protein binding and protein production. However, the protocol and technical information of transient gene expression, especially double strand RNA (dsRNA), in tobacco using Agrobacterium is not well known. Recently, dsRNA is crucial for insecticidal effect on destructive agronomic pest such as Corn rootworm. In this study, we investigated the factor influencing the dsRNA expression efficiency of syringe agro-infiltration in tobacco. To search the best combination for dsRNA transient expression in tobacco, applied two Agrobacterium cell lines and three plant vector systems. The efficiency of dsRNA expression has estimated by real-time PCR and digital PCR. As a result, pHellsgate12 vector constructs showed the most effective accumulation of dsRNA in the cell. These results indicated that the efficiency of dsRNA expression was depending on the kind of vector rather than Agrobacterium cells. In summary, the optimized combination of transient dsRNA expression system in tobacco might be useful to in vivo dsRNA expression for functional study and risk assessment of dsRNA.

국내 LMO 자연환경 모니터링을 위한 11개 LM 옥수수의 동시검출기법 개발

신수영,임혜송,설민아,정영준,김일룡,송해룡,이중로,최원균,Shin, Su Young,Lim, Hae-Song,Seol, Min-A,Jung, Young Jun,Kim, Il Ryong,Song, Hae Ryoung,Lee, Jung Ro,Choi, Wonkyun 한국식물생명공학회 2016 식물생명공학회지 Vol.43 No.4

유전자변형 옥수수의 개발과 상업적 이용이 증가하면서 유전자변형 옥수수 의심시료의 LMO 이벤트를 확인 할 수 있는 적합한 방법 개발이 필요하다. 국내에서는 상업적 재배와 자연생태계의 의도적 비의도적 LMO의 유출이 허용되고 있지 않다. 본 연구에서는 국내 승인된 11개의 LM 옥수수 이벤트를 동시에 검출할 수 있는 동시검출기법을 개발하였다. 이 방법은 시간과 비용, 노동력을 절감할 수 있는 효율적인 방법으로 4종의 PCR set로 개발하였다. 2012년부터 2014년까지 국립환경과학원 및 국립생태원에서 실시한 LMO 자연환경 모니터링 의심시료를 이용하여 동시검출기법의 효율성을 검증하였다. 이러한 결과를 바탕으로 본 연구의 결과는 LMO 모니터링 시료의 분석에 적합한 방법임을 알 수 있었다. With the increasing development and commercial use of genetically modified maize, it is essential to develop an appropriate method for detection of individual LMO (Living modified organism) events for monitoring the samples. In South Korea, commercial planting and accidental or unintentional releases of LMOs into the environment were not approved. In this study, to increase the efficiency of LMO detection, we developed simultaneous detection methods for 11 LM maize events. This multiplex PCR detection method is economical, as it saves time, cost and labor. We developed 11 individual LM maize events, and applied 4 multiplex PCR sets to the LM maize samples. These results are confirmed by applying the multiplex analysis of LMO environmental monitoring from 2012 to 2014, which represents the unintentionally released LM maize samples. The data were correlated with event specific PCR results. Our results indicate that the multiplex PCR method developed is suitable for detection of LM maize in LMO monitoring.

Myeloid deletion of SIRT1 suppresses collagen-induced arthritis in mice by modulatingdendritic cell maturation

우성지,이상명,임혜송,하영술,정인덕,박영민,김현옥,천윤홍,전민규,장규윤,김경민,박병현,이상일 생화학분자생물학회 2016 Experimental and molecular medicine Vol.48 No.-

The type III histone deacetylase silent information regulator 1 (SIRT1) is an enzyme that is critical for the modulation of immune and inflammatory responses. However, the data on its role in rheumatoid arthritis (RA) are limited and controversial. To better understand how SIRT1 regulates adaptive immune responses in RA, we evaluated collagen-induced arthritis (CIA) in myeloid cell-specific SIRT1 knockout (mSIRT1 KO) and wild-type (WT) mice. Arthritis severity was gauged on the basis of clinical, radiographic and pathologic scores. Compared with their WT counterparts, the mSIRT1 KO mice exhibited less severe arthritis, which was less destructive to the joints. The expression levels of inflammatory cytokines, matrix metalloproteinases and ROR-γT were also reduced in the mSIRT1 KO mice compared with the WT mice and were paralleled by reductions in the numbers of Th1 and Th17 cells and CD80- or CD86-positive dendritic cells (DCs). In addition, impaired DC maturation and decreases in the Th1/Th17 immune response were observed in the mSIRT1 KO mice. T-cell proliferation was also investigated in co-cultures with antigen-pulsed DCs. In the co-cultures, the DCs from the mSIRT1 KO mice showed decreases in T-cell proliferation and the Th1/Th17 immune response. In this study, myeloid cell-specific deletion of SIRT1 appeared to suppress CIA by modulating DC maturation. Thus, a careful investigation of DC-specific SIRT1 downregulation is needed to gauge the therapeutic utility of agents targeting SIRT1 in RA.

Event‑specific multiplex PCR method for four genetically modified cotton varieties, and its application

엄순재,김일룡,임혜송,이정로,최원균 한국응용생명화학회 2019 Applied Biological Chemistry (Appl Biol Chem) Vol.62 No.5

Multiplex polymerase chain reaction (PCR) methods have been developed and validated for screening, tracing, and regulating genetically modified (GM) crops in quarantine and environmental monitoring. In this study, we aimed to develop a method to simultaneously detect four GM cotton varieties in order to establish a screening system for cotton volunteers. Based on the sequence of DNA in the junction between introduced gene and flanking genomic DNA of four GM cotton events, herbicide-tolerant MON88701 and DAS-81910-7 and insect-resistant COT102 and T304-40, event-specific primers were designed and a multiplex detection method was developed. The simplex PCR results supported the multiplex PCR results; the amplification efficiency of the novel multiplex PCR method was increased compared with that of the Joint Research Centre (JRC) method. Based on the accuracy and efficiency, the method can be applied to detect and identify randomly mixed reference materials and suspected cotton volunteers. To apply this multiplex PCR method to living modified (LM) environmental monitoring samples, we performed additional PCR analysis to identify whether the volunteers were the four LM cotton varieties. As a result, 66 cotton volunteers were identified with stack event, comprising one or two of the four LM cotton events, and all stacks have been approved in South Korea for food, feed, and processing. These results indicated that our novel multiplex method is suitable for LMO identification.

한국 일러스트레이션 교육의 지평탐색

장미경(Jang Mee Kyung),임혜송(Rim Hye Song) 한국일러스아트학회 1999 일러스트레이션학 연구 Vol.4 No.-

Design has been recently expended in its domain as its field with the great attention of society but Korea illustration has been recognised as supportive art in visual design Field only gaining richness form shallowness and part of decoration. Especially, the education in most universities was not effective both in learning visual esthetic constructive principles and opening eyes to new vision of illustration in this study, I investigated the background story of Korea illustration and problems in university education.